Medical imaging in thick tissues using spatially and temporally modulated light

Bruce J. Tromberg, Ph.D.  

Laser Microbeam and Medical Program
Beckman Laser Institute and Medical Clinic
University of California, Irvine, USA
 

bjtrombe@uci.edu
www.bli.uci.edu

Abstract. This lecture reviews principles of “diffuse optical spectroscopic imaging (DOSI)” for non-invasively characterizing cellular metabolism, extracellular matrix composition, and vascular dynamics in thick tissues. Emphasis is placed on the development of broadband spatially- and temporally-resolved measurements of NIR absorption and scattering spectra. These data are used to form images of deoxygenated hemoglobin, oxygenated hemoglobin, methemoglobin, lipid, and water, as well as the tissue “scatter power”. Clinical study results will be shown highlighting DOSI sensitivity to breast tumor metabolism with sufficient sensitivity for cancer detection and therapeutic drug monitoring. Broadband spatial frequency-domain imaging is used in pre-clinical animal models to dynamically map intrinsic brain signals and monitor the efficacy of chemotherapeutic agents. These findings will be placed in the context of conventional imaging methods, such as MRI, in order to assess the current and future role of diffuse optics in medical imaging.

Biography. Dr. Tromberg is the Director of the Beckman Laser Institute and Medical Clinic (BLI) at the University of California, Irvine. He is a Professor in the departments of Biomedical Engineering and Surgery and has been a member of the BLI faculty since 1990. Dr. Tromberg is principal investigator of the Laser Microbeam and Medical Program (LAMMP), a National Institutes of Health National Biomedical Technology Center. His research interests include the development and application of optical imaging and spectroscopy technologies for non- and minimally-invasive imaging in biology and medicine. He has more than 300 publications and patents in the field of Biomedical Optics and Biophotonics and is a Fellow of the International Society for Optical Engineering (SPIE) and the American Institute for Medical and Biological Engineers (AIMBE).

Photoacoustic tomography: High-resolution in vivo imaging of optical contrast at new depths

Lihong V. Wang, Ph.D.

Gene K. Beare Distinguished Professor

Department of Biomedical Engineering and Department of Radiology

Washington University in St. Louis, USA

lhwang@biomed.wustl.edu

http://oilab.seas.wustl.edu

Abstract. Photoacoustic tomography, as a single hybrid modality, combines the advantages of optical contrast and ultrasonic resolution. High-resolution functional and molecular imaging has been demonstrated in vivo in small animals, while functional imaging has been tested in humans.

Biography. Dr. Wang studied for his Ph.D. degree at Rice University, Houston, Texas under the tutelage of Drs. Robert Curl, Richard Smalley and Frank Tittel. He currently holds the Gene K. Beare Distinguished Professorship in the Department of Biomedical Engineering at Washington University in St. Louis. He has authored and co-authored two books, including one of the first textbooks in the field of biomedical optics. He is the editor for the first comprehensive book on biomedical photoacoustic tomography. He has published 195 peer-reviewed journal articles and delivered 216 keynote, plenary, and invited talks. He is a fellow of the American Institute for Medical and Biological Engineering, the Optical Society of America, the Institute of Electrical and Electronics Engineers, and the Society of Photo-Optical Instrumentation Engineers. He was appointed as the Editor-in-Chief of the Journal of Biomedical Optics. He serves as an equal co-chair for the annual conference on Photons plus Ultrasound, the 2010 Gordon Conference on Lasers in Medicine and Biology, and the 2010 OSA Topical Meeting on Biomedical Optics. He also serves as an equal co-chair for the International Biomedical Optics Society. He has served as a study section chair or grant reviewer for NIH and NSF. He is currently a chartered member on an NIH study section. He serves as the founding chair for the scientific advisory board of a company commercializing his invention. His research on non-ionizing biophotonic imaging has been funded with a cumulative budget of >$25M (principal investigator for 21 research grants) by NIH, NSF, and other funding agencies. He was a recipient of the NIH FIRST award and NSF CAREER award.

His laboratory invented or discovered frequency-swept ultrasound-modulated optical tomography, dark-field confocal photoacoustic microscopy (PAM), optical-resolution PAM, photoacoustic Doppler sensing, photoacoustic reporter gene imaging, focused scanning microwave-induced thermoacoustic tomography, exact reconstruction algorithms for photoacoustic or thermoacoustic tomography, sonoluminescence tomography, Mueller-matrix optical coherence tomography, optical coherence computed tomography, and oblique-incidence reflectometry. In particular, PAM broke through the long-standing penetration limit of conventional optical microscopy due to photon diffusion and reached super-depths for noninvasive biochemical, functional, and molecular imaging in living tissue at high resolution. His Monte Carlo model of photon transport in scattering media has been used worldwide

Raman microspectroscopy –  a powerful tool for biomedical diagnosis

Institut für Physikalische Chemie
Friedrich-Schiller-Universität Jena
Jena, Germany

juergen.popp@ipht-jena.de

www.ipc.uni-jena.de

Abstract. Here we will present challenges to be met in connection with the application of molecular spectroscopic imaging and in particular Raman microspectroscopy for life sciences and biomedicine. Overall within this contribution it will be shown that Raman microspectroscopy and its various techniques (micro-Raman, SERS, CARS, TERS etc.) are powerful biophotonic tools for bioanalytical and biomedical applications like e.g. rapid pathogen identification, sensitive drug monitoring or clinical tissue diagnostics.

Biography. Dr. Popp was born in 1966, received his Ph.D. in chemistry from the University of Würzburg, Germany, in 1995. In 1996 he spent a year as a visiting scientist in the Department of Applied Physics of Yale University, New Haven, USA. He subsequently joined the group of Prof. Dr. W. Kiefer in the Institute of Physical Chemistry, University of Würzburg where he finished his habilitation in 2000. Since May 2002 he is a full professor at the Friedrich--Schiller university of Jena, Germany where he holds a chair of physical chemistry. In June 2006 he also became the Scientific Director at the Institute of Photonic Technology. His work has been awarded by the faculty prize of chemistry (1995), by the “Bayerischer Habilitationsförderpreis” (1997), by the “Förderpreis der Würzburger Korporationen” (2001) and the Kirchhoff--Bunsen award (2002). Since 2009 he is a fellow of the Society for Applied Spectroscopy.

The research interests of J. Popp are mainly centered around the development and application of frequency-, time- and spatially resolved innovative laser spectroscopical methods and techniques ranging from the UV into the NIR region for the derivation of structure activity or dynamic relationships. This type of investigative approach is essential in resolving important questions in fields of biology, medicine, pharmacy, astronomy as well as in the environmental and material sciences. In particular his expertise is in the field of Raman spectroscopy and in the development of innovative Raman techniques should be emphasized. The results obtained by J. Popp were published in more than 200 scientific articles in premier peer-reviewed journals. He is inventor of 5 patents in the field of spectroscopical instrumentation. J. Popp is editor in chief of Journal of Biophotonics and editorial board member of Journal of Raman spectroscopy and ChemPhysChem.

Keynote and invited speakers

Smart nanoparticles for imaging and drug delivery, keynote paper

University of Washington (USA)

Biography. Prof. Xiaohu Gao received his Ph.D. degree in chemistry from Indiana University, Bloomington in 2004, and his postdoctoral training from the Department of Biomedical Engineering at Georgia Tech and Emory University. He became a faculty member in the Department of Bioengineering and the Center for Nanotechnology at the University of Washington, Seattle in 2005. His research is focused on biomedical nanotechnology, molecular engineering and optical imaging. He has authored or co-authored over 40 papers and book chapters; and he is also a recipient of the NSF CAREER Award. He has been a member of the American Chemical Society (ACS) and Biomedical Engineering Society (BMES) since 2003.

Abstract. Application of nanotechnology and photonics in medicine has the potential to transform disease diagnostics and treatment. In this talk, I present recent development of multifunctional nanoparticles for molecular imaging and controlled delivery of therapeutic drugs.

Invited speakers

Saulius Bagdonas, Vilnius University, Lithuania, "Phototransformations of quantum dots: intersection of coating, environment and light"

Kristian Berg, The Norwegian Radium Hospital, Norway, "From bench to bedside with a novel technology for site-specific drug delivery"
Sergey Deyev, Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russia, "Protein self-assembler for synthesis of multifunctional heterostructures"

Alexander Priezzhev, M.V. Lomonosov Moscow State University, Russia, "Laser assessment of the effect of diamond nanoparticles on deformability and aggregation of red blood cells in vitro"

Ricardas Rotomskis, Vilnius University, Lithuania, "Nanoparticles in photodynamic therapy of tumour"

Andrey Zvyagin, Macquarie University, Australia, "Interfacing nanodiamonds with the biomolecule world"

2. Laser-Tissue Interactions

Morpho-functional nonlinear laser imaging of tissues, keynote paper

Francesco Pavone

Universita degli Studi di Firenze (Italy)

Biography. Francesco Saverio Pavone was born in Bari the 23^th March 1962. In 1989 he obtained his Laurea degree in Physics at the University of Florence. In 1990 he became Research Officer at the European Laboratory for Non-Linear Spectroscopy (University of Florence). In 1993 he obtained  a Ph.D. in Optics at the National Institute for Optics (Florence, Italy). In 1997 he spent one year and half as "Maitre de Conférences Associe au College de France", Paris, with experimental work at  the "Ecole Normale Superieure" (ENS) of Paris with Prof. Claude Cohen-Tannoudji (199? Nobel price in Physics). In 1998 he became associate Professor of physics at the department of physics of the University of Perugia, Italy and Scientific director of the section of Atomic and Molecular Physics at the European Laboratory for Non-Linear Spectroscopy (LENS), Florence, Italy. In 2001 he moved as associate professor to the university of Florence (dept. of physics) and became scientific responsible of the “biophysics laboratory” of the European Laboratory for Non-Linear Spectroscopy (Florence, Italy). In 2005  he became full professor. As scientific experience, from 1990 to 1995 he worked  in the field of “Atomic and molecular spectroscopy”. From 1995  to 1999 he worked in the field of Atomic physics, and since 1999 in Biophysics. Currently he is directing a research group working in the field of biophotonics on single molecule biophysics, microscopy imaging-spectroscopy techniques, biomedical imaging, laser manipulation of bio-samples. In particular, he is developing new microscopy techniques for high resolution and high sensitivity imaging, and for laser manipulation purposes. These techniques have been applied both for single molecule biophysics, single cell imaging and optical manipulation. Tissue imaging is an other research area developed, where non linear optical techniques have been applied for sking and neural tissue imaging. Recently, In-Vivo imaging apparatus have been developed and applied to animal and humans. Pavone is authors of many internationals papers and editor of international books. He has more than 50 invited talk and he is editors of international journals. He coordinates several European projects and he has organized international congresses; he is also director of the international PhD program at LENS.

Abstract. We will present some non linear imaging applications, where Multiphoton, SLIM, FLIM or SHG imaging are used. Morphological and functional aspects of the tissue will be investigated. Applications to neurological tissues, skin, bladder or cornea will be shown. Morphological features, also following laser damaging, or functional aspect related to signal propagation or cell signaling in tissues will be described in the paper.

Invited speakers

Steven Jacques, Oregon Health and Science University, USA, "How tissue optical properties affect dosimetry for laser procedures and phototherapy"

Alexander Krasnovsky, A.N. Bach Institute of Biochemistry, Moscow, Russia, "Summation of energy of two singlet oxygen molecules in dye containing systems under laser excitation. Mechanisms and analytical applications"

Horacio Lamela, Carlos III de Madrid University, Spain, "Novel interferometric sensors for optoacoustic imaging and biomedical applications"

Kirill Larin, University of Houston, USA, "Noninvasive functional imaging of early embryonic development in mammalian systems with Optical Coherence Tomography"

Victor Loschenov, A.M. Prokhorov General Physics Institute, Russia, "Structural transformation of nanophotosensitizers in biological environments"

Elena Zagaynova, Nizhny Novgorod State Medical Academy, Russia, "Optical diagnostics and laser hyperthermia of tumors with plasmon-resonance gold nanoparticles"

3. Laser Biomedical Diagnostics, Sensing and Therapy

Using elastic scattering spectroscopy to reveal early stages of apoptosis in viable cells, keynote paper

Medicine Boston University (USA)

Biography. Irving J. Bigio received his Ph.D. in Physics from the University of Michigan in 1974. From then until 2000 he was a scientific staff member at Los Alamos National Laboratory (New Mexico), including service as Leader of the Laser Science and Applications Program (1988-1994). During various leaves of absence he was a Fulbright Senior Scholar at the Weizmann Institute of Science, in Israel, a Visiting Professor at the University of Copenhagen, Denmark and a Guest Fellow of Pembroke College at the University of Oxford, England. Dr. Bigio is inventor on a number of patents for biomedical optics instrumentation, and has received three R&D-100 Awards for the development of biomedical optical devices.  Since February 2001 he has been Professor at Boston University, in the Departments of Biomedical Engineering, Electrical & Computer Engineering, Physics, and Medicine. He is also Honorary Guest Professor of the University College London, Department of Surgery. Dr. Bigio serves on several government advisory panels and on external advisory boards for companies and academic institutions. He is a Fellow of the Optical Society of America and the American Institute of Medical and Biological Engineering, and is a member of the American Physical Society and the SPIE. In addition to other research projects in biomedical optics, Dr. Bigio recently led a multi-institutional program under the NIH/NCI Network for Translational Research in Optical Imaging, comprising several medical research centers in the US and Europe. 

Abstract. Apoptosis, “programmed cell death,” is a cellular process exhibiting distinct biochemical and morphological changes. It is one of the most studied phenomena in cellular biology, pathology and medicine in general. There are journals dedicated to apoptosis, and thousands of papers are published annually dealing with the process, the factors that induce or inhibit it, and its role in many diseases, especially cancer. With very few exceptions, scientists studying apoptosis in cell cultures perform standardized assays that are invasive and labor intensive. They require fixing the cells at specific time points and tagging them with stains or fluorophores, followed by microscopic examination or flow cytometry. Typically the earliest signs of apoptosis are seen 1-2 hours following exposure or treatment by an inducing agent. Any earlier indications are below the resolution limits of optical microscopy, and are not yet evidenced by membrane disruption (which is used for tagging for flow cytometry). A user-friendly method to track apoptotic changes in viable cells, and at earlier time points, would be of great value to the community. We have developed a noninvasive method based on light backscattering spectroscopy, which constitutes the first report of a noninvasive method to monitor apoptosis that is sensitive to changes that occur as early as 10 minutes after treatment. Cristine’s method is noninvasive: i.e., the measurements do not require disruption of the cell culture and can be used continuously to monitor cell status at any time point.  Moreover, the method is quantitative, providing information about the size distribution of subcellular structures that are active in the apoptotic process, thus providing new information on the apoptotic pathways. These include structures as small as 50 nm or less, much smaller than the resolution limit of optical microscopy. We believe this novel optical method can become a valuable tool for research in cellular biology and disease pathology on the microscopic scale.

Invited speakers

Stefan Andersson-Engels, Lund University, Sweden, "Diffuse fluorescence spectroscopy for tissue diagnostics and treatment control"

Rafat Ansari, NASA Glenn Research Center, USA, "Non-invasive and early detection of oxidative stress leading to normal aging on Earth and accelerated aging during spaceflight"

Darryl Bornhop, Vanderbilt University, USA, "Molecular interaction studies on membrane-bound proteins using backscattering interferometry"

Juergen Lademann,  Charité – Universitätsmedizin Berlin, Germany, "Interaction between antioxidants and free radicals in human skin"

Martin Leahy, Univ. of Limerick, Ireland, "Challenges in deep tissue imaging"
Galina Petrova, M.V. Lomonosov Moscow State University, Russia, "Physical mechanism of poisoning proteins and enzymes by heavy metals"

Valery Tuchin, Saratov State University, Russia, "Laser cytometry in vivo"

4. Single Cells and Molecules; Optical Trapping and Manipulation

Feeling for cells with light: Illuminating the role of biomechanics for tumor progression,

keynote paper

University of Leipzig (Germany)

Abstract. Light has been used to observe cells since Leeuwenhoek’s times; however, we use the forces caused by light described by Maxwell’s surface tensor to feel for the cellular cytoskeleton.  The cytoskeleton, a compound of highly dynamic polymers and active nano-elements inside biological cells, is responsible for a cell’s stability and organization.  The optical stretcher exploits the nonlinear, thus amplified response of a cell’s mechanical strength to small changes between different cytoskeletal proteomic compositions as a high precision cell marker that uniquely characterizes different cell types.  Consequentially, the optical stretcher detects tumors and their stages with accuracy unparalleled by molecular biology.  As implied by developmental biology the compartmentalization of cells and the epithelial-mesenchymal transition that allows cells to overcome compartmental boundaries strongly depend on cell stiffness and adhesiveness. Consequentially, biomechanical changes are key when metastatic cells become able to leave the boundaries of the primary tumor.

Invited speakers
Kishan Dholakia, University of St Andrews, Scotland, "Light takes shape: Advanced biophotonics with spatial light modulation"

Jesper Glückstad, Technical University of Denmark, Denmark, "Next generation of biophotonics workstation"

Mathias Goksör, University of Gothenburg, Sweden, "Single cell analysis using optical manipulation"

Karl Otto Greulich, Leibnitz Institute for Age Research, Germany, "DNA damages induced by UV VIS (-lasers) and their repair in pharmacological therapy and ageing"

Karin Schütze, CellTool GmbH, Germany, "From scientific results to marketable products"

5. THz Waves in Biophotonics

THz technique for skin measurement, keynote paper

Biography. Kodo Kawase was born in Nagoya in 1966. He received the B.S. degree in Electrical Engineering (EE) from Kyoto Univ. in 1989, and the Ph. D degree in EE from Tohoku Univ. in 1996. He became a Unit Leader at RIKEN in 2001. He became a professor of Agricultural Science, Tohoku Univ. in 2004. In July 2005, he became a Professor of EE at Nagoya Univ. He also became a Leader of Terahertz Application Team at RIKEN in 2008. He has received the 2005 Young Scientists’ Prize by the Commendation for Science and Technology by the Minister of Education, and six other prizes

Abstract. Our group has been conducting research activities in several directions within the THz field. We introduced many types of widely tunable THz-wave sources using nonlinear optical effects, and we also suggested a whole range of real-life applications using THz-imaging techniques. Recently, we are developing several novel THz techniques for skin measurement.

The THz dance of the protein with the water, keynote paper

Biography. Prof. Dr. Martina Havenith-Newen, studied physics in Bonn (diploma 1987). She received several grants such as the oversea grant (UC Berkeley 1987-88) from the German National Merit Foundation, a Habilitation grant (1995-98) and the Heisenberg grant (1998) of the German Science Foundation. Since 1998 she holds a chair of physical chemistry in Bochum. In 1995 she received the Bennigsen-Förder-Award of the State of North Rhine-Westphalia and in 2004 the Human Frontier Science Programme Award.  She is a full member of the North Rhine-Westphalia Academy of Science and a member of the German Academy of Science, the Leopoldina.

Abstract. The focus in protein folding has been very much on the protein backbone and sidechains.  Yet hydration waters make comparable contributions to the structure and energy of proteins. Although the dynamics of the hydration water occurs on the picosecond time scale, ‘slaving’ to fast solvent modes profoundly affects the slower but larger-scale protein motions. In return the protein influences the structure and dynamics of surrounding water molecules. Fundamental questions of biomolecule hydration include, how far out into the solvent does the influence of the biomolecule reach, how is the water affected, and how are the properties of the hydration water influenced by the separation between protein molecules in solution? Terahertz spectroscopy is shown to directly probe such solvation dynamics, and the width of the dynamic hydration layer. For the proteins such as the five helix bundle protein l_6-85 we found dynamical hydration layers which extend over more than 20 Å. This is greater than the static structural correlation length observed by X-ray spectroscopy. Most recent results demonstrate that the long range hydration layer is protein sequence and pH dependent. It is interesting to note that the native wildtype shows the most pronounced effect on the fast water dynamics. The denaturated protein and any mutants have much less influence on the fast water network motions. We could further support the idea that the protein dynamics is slaved to the solvent dynamics by time resolved studies. We have introduced Kinetic Terahertz Absorption spectroscopy (KITA) and could show that the rearrangement of the protein-water network motions as probed by Terahertz spectroscopy upon is participating in the initial steps during protein folding. We have recently extended our studies to small peptides which showed that the required hydration level for the observation of an onset of collective network modes in the THz range corresponds to the number of water molecules which are required for biological functionality.

Invited speakers

Robin Bocquet, University de Littoral Cote d'Opale, France, "The need of high accuracy frequency measurements for detection of atmospheric pollutants"

David Plusquellic, National Institute of Standards and Technology, USA, "State-resolved THz spectroscopy and dynamics of peptide/water systems"

Alexander Shkurinov, M.V. Lomonosov Moscow State University, Russia, "THz time-domain spectroscopy and spectrochromography from basis to spectral line assignment"

Joo-Hiuk Son, University of  Seoul, Korea, "Terahertz molecular imaging for medical applications"
Ingrid Wilke, Rensselaer Polytechnic Institute, USA, "THz emitter development for THz wave application in biophotonics"

Gerald Wilmink, Air Force Research Laboratory, Brooks City-Base, USA, "Assessing interactions between Terahertz radiation and biological tissue, cellular, and molecular level"

6. Vibrational Spectroscopy of Biological Systems

Development of UV Raman spectroscopy for incisive investigations of simple questions in complex systems: Can we study the reaction coordinate in protein folding?, keynote paper

University of Pittsburg (USA)

Biography. Sanford A. Asher, Distinguished Professor of Chemistry at the University of Pittsburgh received his B.A. in chemistry at the University of Missouri, St. Louis in 1971 and Ph.D. in chemistry at the University of California, Berkeley in 1977.  Dr. Asher was a Research Fellow in Applied Physics at Harvard University between 1977 and 1980.  In 1980 he became Assistant Professor of Chemistry at the University of Pittsburgh.  Dr. Asher’s research program at Pitt has involved development of new materials and development of new spectroscopic techniques.  His group developed UV resonance Raman spectroscopy as a new technique for fundamental and applied structural and trace studies of molecules in complex matrices.  His group is using UV resonance Raman to examine the first stages in protein folding.  In addition, Dr. Asher’s research group develops new photonic crystal optical devices and chemical sensing devices from self-assembling colloidal particles.  He has pioneered the development of smart hydrogel materials.

Abstract. UV Raman excitation into these ~200 nm electronic transitions results in the enhancement of the amide vibrations of the peptide backbone.   In our most recent studies we reassigned the amide III region and found a particular band (the amide III₃ band) which reports selectively on the Ramachandran Ψ angle and the state of peptide bond hydrogen bonding.  We demonstrate that this band is Raman scattered independently by each peptide bond with insignificant coupling between peptide bonds.  We also show that isotope editing of a peptide bond (by replacing the C_α - H with C_α - D) allows us to determine the frequency of an individual peptide bond within a peptide or protein which gives us its Ψ angle.  Consideration of the Boltzmann equilibria allows us to determine the Ψ angle energy landscape which connects secondary structure conformations.  The Ψ angle coordinate is the most important reaction coordinate required to enable the understanding of the mechanism(s) of protein folding.

Invited speakers

Andrey Chikishev, M.V. Lomonosov Moscow State University, Russia, "Laser spectroscopy and computer simulation of the effect of solvent molecules on protein dynamics and function"

Hideaki Kano, University of Tokyo, Japan, "Label-free, multi-color, high-speed imaging of a living cell by CARS spectral imaging"

Jouko Korppi-Tommola, University of Jyväskylä, Finland, "Modelling excitation transport in photosynthetic light harvesting complexes"

Hristo Iglev, Technische Universität München, Germany, "Electron detachment and recombination in aqueous halides studied with 2- and 3-pulse femtosecond spectroscopy"

Leonard Proniewicz, Jagiellonian University, Poland, "Surface enhanced Raman spectroscopy (SERS) of selected neurotransmittersbombesin family compounds, analogs and fragments"

Cees Otto, University of Twente, The Netherlands, "Spontaneous Raman microscopy and CARS microscopy of developing bone minerals in in vitro tissues"

Hiroaki Takahashi, Waseda University, Japan, "Structure and dynamics of electronic excited states of dibenzazepine"

Wolfgang Werncke, Max-Born-Institut, Germany, "Vibrational energy redistribution after NH-stretching excitation in hydrogen-bonded dimers and DNA oligomers"

7. Molecular and Bio-Imaging

Illuminating biomedical discovery with multi-spectral opto-acoustic tomography (MSOT), keynote paper

Technische Universität München (Germany)

Biography. Vasilis Ntziachristos, Ph.D. is a Professor and Chair for Biological Imaging at the Technische Universtität München and the Helmholtz Zentrum München and the director of the Institute for Biological and Medical Imaging. Prior to this appointment he has been faculty at Harvard University and the Massachusetts General Hospital. He has received his masters and doctorate degrees from the Bioengineering Department of the University of Pennsylvania and the Diploma on Electrical Engineering from the Aristotle University of Thessaloniki, Greece. Professor Ntziachristos serves as chair in international meetings and in the editorial boards of scientific journals and was named one of the world’s top innovators by the Massachusetts Institute of Technology (MIT) Technology Review in 2004. His main research interests involve the development of optical methodologies for probing physiological and molecular events in tissues using non-invasive methods.

Abstract. Optical imaging is unequivocally the most versatile and widely used visualization modality in clinical practice and life sciences research. In recent years, advances in photonic technologies and image formation methods have received particular attention in biological research and the drug discovery process for non-invasively revealing information on the molecular basis of disease and treatment. An increasing availability of endogenous reporters such as fluorescent proteins and probes with physiological and molecular specificity enable insights to cellular and sub-cellular processes through entire small animals, embryos, fish and insects and have revolutionized the role of imaging on the laboratory bench, well beyond the capability of conventional microscopy. This talk describes current progress with instruments and methods for in-vivo photonic tomography of whole intact animals and model biological organisms. We show how new tomographic concepts using opto-acoustics are necessary for accurate and high-resolution quantitative molecular investigations in tissues and why it could be potentially a valuable tool for accelerated investigations of therapeutic efficacy and outcome. We further demonstrate that cellular function and bio-chemical changes can be detected in-vivo, through intact tissues at high sensitivity and molecular specificity. Examples of imaging enzyme up-regulation, carcinogenesis and gene-expression are given. The potential for clinical translation is further outlined. Limitations of the method and future directions are also discussed.

Invited speakers

Regine Choe, University of Pennsylvania, USA, "In vivo cancer therapy monitoring with diffuse optics"

Frank Chuang, University of California, Davis, USA, "Biophotonic applications in molecular medicine"

Hamid Dehghani, University of Birmingham, UK, "Current developments and challenges in molecular diffuse optical and bioluminescence tomography"

Qingming Luo, Huazhong University of Science and Technology, China, "NIR imaging of human prefrontal cortex activity for verbal n-back tasks"

Ilya Turchin, Institute of Applied Physics, Russia, "Fluorescence tomography of red fluorescent protein expressed tumors in small animals"

8. Laser Microscopies

Multi-dimensional microscopy of living cells, keynote paper

Biography. Herbert Schneckenburger is a professor of Physics, Optics and Biophotonics at Hochschule Aalen since 1986 and a private lecturer of the Medical Faculty of the University of Ulm since 1992. After studying Physics in Stuttgart and Grenoble he passed his PhD at the University of Stuttgart in 1979 and his habilitation at the University of Ulm in 1992. From 1979-1986 he built up a laboratory for time-resolved laser spectroscopy at the Helmholtz-Zentrum (formerly GSF Research Centre) in Munich. Prof. Schneckenburger’s present interests are focused on novel methods and applications of fluorescence microscopy, laser spectroscopy and biomedical screening.

Abstract. Methods of laser-assisted fluorescence microscopy with high spatial, spectral and temporal resolution are combined and used for diagnostics of living cells under controlled light exposure. An overview on techniques and recent applications of laser-assisted fluorescence microscopy with high spatial, spectral and temporal resolution is given. Spectral imaging is used to describe various properties of living cells, e.g. membrane stiffness as a function of temperature and cholesterol content. Fluorescence lifetime seems to be an appropriate parameter to assess malignancy in tumour cells, but can also be used to probe molecular interactions via Förster Resonance Energy Transfer (FRET), e.g. in studies of Alzheimer’s disease or in sensing of apoptosis. Variable-angle total internal reflection fluorescence microscopy (VA-TIRFM) as well as methods of structured illumination are used to obtain high axial resolution, whereas polarization microscopy is applied for measuring cell and membrane dynamics. It should be emphasized that present experiments are performed under controlled light exposure in order to avoid damage to living cells, even if resolution and sensitivity may thus be limited. In addition, it appears important to introduce 3-dimensional cellular systems, which may describe tissue properties better than a cell monolayer (e.g. growing on a glass slide). Marker free microscopy (including autofluorescence and Raman microscopy) is of increasing interest in view of future in vivo experiments. Finally, fluorescence microscopy is expected to stimulate further techniques, e.g. fluorescence screening or laser-assisted micromanipulation.

Invited speakers

Chia-Liang Cheng, National Dong Hwa University, Taiwan, "Nanodiamond interaction with human red blood cells: The microspectroscopic point of view"

Alzbeta Chorvatova, International Laser Centre, Slovakia, “Time-resolved micro-spectroscopy of endogenous metabolites in living cells”

Alexander Savitsky, A.N. Bach Institute of Biochemistry, Russia, "Fluorescence life-time imaging of enzyme activities in living cells"

Timo Zimmermann, Centre for Genomic Regulation, Spain, "Fluorophore illumination effects in different laser scanning microscopy methods: A comparison of single-beam and multi-beam confocal microscopy and multiphoton microscopy method"

9. Novel Optical Devices for Biomedicine

Optical imaging in ophthalmology, keynote paper

University of Tsukuba (Japan)

Biography. Dr. Yoshiaki Yasuno is an assistant professor of University of Tsukuba and directs Computational Optics Group in the University of Tsukuba since 2002. In this group, he researches a high-speed ophthalmic optical coherence tomography (OCT), polarization sensitive OCT, Doppler OCT and its applications to ophthalmology and dermatology.  He received his PhD of Engineering in 2001 from University of Tsukuba for his work of spatio-temporal optical computing.

Abstract. Since eye is only one optical organ in a human body, there are several optical-testing modalities of the eye. In this talk I would like to give a review of the optical modalities in the ophthalmology.

Invited speakers

Barry Cense, Utsunomiya University, Japan, “Measuring polarization properties of the human retina with polarization-sensitive OCT and adaptive optics”

Arthur Chiou, National Yang-Ming University, Taiwan, "Probing the viscoelastic properties of individual human RBCs by optical trap-and-stretch; a brief overview and recent progresses"

Karl-Heinz Feller, University of Applied Sciences Jena, Germany, "Microfluidic lab-on-a-chip platform for bioanalytics and bimolecular photochemistry"

Kirill Linkov, A.M. Prokhorov General Physics Institute, Moscow, Russia, "New trends in medical equipment for photodynamic therapy and fluorescent diagnostics"

Rudolf Steiner, Universität Ulm, Germany, "Spectroscopic online diagnostics for laser therapy"

10. Printing Techniques and their Applications in Biotechnology

To be announced, keynote paper

Ghassan Jabbour

King Abdullah University of Science and Technology (Saudi Arabia)

and

University of Oulu (Finland)

Biography. To be added.

Abstract. To be added.

Invited speakers

Khaled Salama, King Abdullah University of Science and Technology, Saudi Arabia, "Integrated biosensors".

11. Symposium on Water in Bioenvironment: Spectroscopy and Simulation

Surprising effect of light on water, keynote paper

University of Washington (USA)

Biography. Gerald Pollack, Ph.D. received his doctorate in biomedical engineering from the University of Pennsylvania in 1968. He then joined the University of Washington faculty and is now professor of Bioengineering. His interests have ranged broadly, from biological motion and cell biology to the interaction of biological surfaces with aqueous solutions. His 1990 book, Muscles and Molecules: Uncovering the Principles of Biological Motion, won an “Excellence Award” from the Society for Technical Communication; his more recent book, Cells, Gels and the Engines of Life, won that Society’s “Distinguished Award.” Pollack received an honorary doctorate in 2002 from Ural State University in Ekaterinburg, Russia, and was more recently named an Honorary Professor of the Russian Academy of Sciences. He received the Biomedical Engineering Society’s Distinguished Lecturer Award in 2002. More recently, in 2008, he was selected among all University of Washington faculty to receive the faculty’s highest distinction: the Annual Faculty Lecturer Award. Pollack is a Founding Fellow of the American Institute of Medical and Biological Engineering and a Fellow of both the American Heart Association and the Biomedical Engineering Society. He is also Founding Editor-in-Chief of the new journal, WATER, and has recently received an NIH Transformative R01 Award.

Abstract. The impact of surfaces on the contiguous aqueous phase is generally thought to extend no more than a few water-molecule layers. We find, however, that colloidal and molecular solutes are profoundly excluded from the vicinity of hydrophilic surfaces, to distances typically several hundred micrometers. Such large zones of exclusion have been observed next to many different hydrophilic surfaces, and many diverse solutes are excluded. Hence, the exclusion phenomenon appears to be quite general. To test whether the physical properties of the exclusion zone differ from those of bulk water, several methods have been applied. NMR, infrared, and birefringence imaging, as well as measurements of electrical potential, viscosity, and UV-VIS and infrared-absorption spectra, collectively reveal that the solute-free zone is a physically distinct, more ordered phase of water. It can co-exist essentially indefinitely with the contiguous solute-containing phase. Indeed, this unexpectedly extensive zone may be a candidate for the long-postulated “fourth phase” of water considered by earlier scientists. The energy responsible for building this charged, low entropy zone comes from light. We found that incident radiant energy including all visible and near-infrared wavelengths induce exclusion-zone growth in a spectrally sensitive manner. IR is particularly effective. Five-minute exposure to radiation at 3.1 µm (corresponding to OH stretch) causes exclusion-zone-width increase up to three times. Apparently, incident photons cause some change in bulk water that predisposes constituent molecules to reorganize and build the charged, ordered exclusion zone. We found also that such photons can power the flow of water through small hydrophilic tubes, with no additional source of energy. Photons from ordinary sunlight, then, may have an unexpectedly powerful effect that goes beyond mere heating. It may be that solar energy builds order and separates charge between the near-surface exclusion zone and the bulk water beyond — the separation effectively creating a battery. The resemblance to the first steps of photosynthesis is evident. Indeed, this light-induced action would seem relevant not only for photosynthesis, but also for all realms of nature and engineering involving water and interfaces, and also for biology, where much or all of the cell’s water may be structured. The implications are amply discussed in http://uwtv.org/programs/displayevent.aspx?rID=22222 and http://www.i-sis.org.uk/liquidCrystallineWater.php and will be presented in the lecture.

Invited speakers

Nikolay Bunkin, A.M. Prokhorov General Physics Institute, Russia, "Stable gas nanobubbles in water and acqueous solutions of salts; their role in living processes and bioenvironment"

Thomas la Cour Jansen, University of Groningen, The Netherlands, "Hydrogen bonding and water dynamics in peptide systems probed by non-linear infrared spectroscopy"

Elmar Fuchs, Wetsus — Centre of Excellence for Sustainable Water Technology, The Netherlands, "Mass and charge transfer within a floating water bridge"

Sergey Pershin, Prokhorov General Physics Institute, Moscow, Russia, "Quantum origin of a jump in erythrocyte penetration through a microcapillary at 36.6 ⁰C: ortho-para H₂O in water"

Maxim Pshenichnikov, University of Groningen, The Netherlands, "Water dynamics near hydrophobes: What can 2D IR teach us?"

Roumiana Tsenkova, Kobe University, Japan, "Aquaphotomics: Water light interaction as biological marker"